Dear Ronnie,
I would suggest to cut off the standard lane!! and process the standard and
the test sample separately in the appropriate way. You can still compare the
two, no problem.
Or - change from the biotinylated standard to a prestained standard (which
is sold by lots of companies)and does not need any further processing, less
work.
regards,
Angelika
-----Ursprüngliche Nachricht-----
Von: Ronnie Houston [mailto:wee_rory@hotmail.com]
Gesendet: Mittwoch, 28. Juni 2000 21:08
An: histonet@pathology.swmed.edu
Betreff: Western Blots
Not strictly a Histo question, but one of our researchers is doing some
Western Blots for us on skeletal muscle samples from suspected Beckers
Muscular Dystrophy.
She is using biotinylated standards. However, the primary dystrophin
antibody is not biotin-conjugated, so she is applying a biotinylated
secondary anti-mouse Ig to both test samples and the standard. This is
followed by an avidin-conjugated alkaline phosphatase and visualization with
BCIP/NBT.
My question is, can you compare the results of the standard to the test when
the biotinylated standard has been subjected with IgG-biotin? Does this
extra amount of biotin affect the result?
My initial thoughts are that one cannot directly compare results because
differently prepared samples have been run. Also there has been no blocking
for endogenous biotin.
Thanks for any help you can offer.
Ronnie Houston
Cytochemistry & Molecular Pathology
Texas Scottish Rite Hospital for Children
2222 Welborn Street
Dallas, TX 75219
(214) 559 7744
(214) 559 7768 - fax
rhouston@tsrh.org
________________________________________________________________________
Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com